Criopreservação de células-tronco mesenquimais do tecido adiposo: Uma alternativa para lipoenxertia seriada na reconstrução mamária / Cryopreservation of adipose tissue-derived mesenchymal stem cells: An alternative to serial fat grafting in breast reconstruction

Introdução: A lipoenxertia é um enxerto autólogo de células do tecido celular subcutâneo, que pode ser utilizada como técnica complementar na reconstrução mamária. Diante disso, a criopreservação de células-tronco mesenquimais provenientes de tecido adiposo (CTDAs) poderia ser uma maneira de realizar a coleta em um tempo cirúrgico e após realizar a lipoenxertia de forma fracionada. O dimetilsulfóxido (DMSO) é um criopreservante utilizado em pesquisas com células, porém é potencialmente tóxico, o que impossibilitaria a utilização de CTDAs criopreservadas na prática clínica. Novos criopreservantes celulares, sem toxicidade, vêm sendo descritos na literatura científica experimental, como as substâncias L-prolina e trealose. Com isso, esse trabalho teve como objetivo avaliar a viabilidade de CTDAs criopreservadas com a combinação de L-prolina e trealose, em um período de até 90 dias. Método: Estudo experimental, no qual foram obtidas amostras de lipoaspirado provenientes de 9 pacientes. A fração celular foi processada e congelada com L-prolina (1,5M) + trealose (0,2M), ou com DMSO + soro fetal bovino (SFB), como controle. Após 30 e 90 dias, as amostras foram descongeladas e a viabilidade celular foi avaliada pela técnica de MTT. Resultados: A análise das CTDAs, após 1 e 3 meses de congelamento, indicou que as amostras tratadas com L-prolina + trealose apresentaram viabilidade semelhante àquelas preservadas com DMSO e SFB (p=0,444). Conclusão: A associação de L-prolina e trealose manteve CTDA viáveis por 30 e 90 dias de congelamento, podendo ser uma alternativa como criopreservante celular sem toxicidade e viabilizando o uso de lipoenxertia seriada.

Introduction: Fat grafting is an autologous graft of cells from subcutaneous tissue, which can be used as a complementary technique in breast reconstruction. Given this, the cryopreservation of adipose tissue-derived mesenchymal stem cells (ADMSCs) could be a way to collect them in one surgical procedure and after performing fractional fat grafting. Dimethyl sulfoxide (DMSO) is a cryopreservative used in cell research, but it is potentially toxic, which would make it impossible to use cryopreserved ADMSCs in clinical practice. New cellular cryopreservatives, without toxicity, have been described in the experimental scientific literature, such as the substances L-proline and trehalose. Therefore, this work aimed to evaluate the viability of ADMSCs cryopreserved with the combination of L-proline and trehalose over up to 90 days. Method: Experimental study in which lipoaspirate samples were obtained from 9 patients. The cellular fraction was processed and frozen with L-proline (1.5M) + trehalose (0.2M) or with DMSO + fetal bovine serum (FBS) as control. After 30 and 90 days, the samples were thawed, and cell viability was assessed using the MTT technique. Results: The analysis of ADMSCs, after 1 and 3 months of freezing, indicated that samples treated with L-proline + trehalose showed similar viability to those preserved with DMSO and SFB (p=0.444). Conclusion: The association of L-proline and trehalose kept ADMSC viable for 30 and 90 days of freezing, and could be an alternative as a cellular cryopreservative without toxicity and enabling the use of serial fat grafting.

High fat diet- induced obesity decreases ErbB receptors expression in liver and adipose tissue in C57BL/6 mice

Neuregulins (NRGs) are a family of signaling proteins that bind to receptor tyrosine kinases of the ErbB family (ErbB2 to ErbB4), which can homo- or heterodimerize depending on their structural features and cell type. Many studies have proposed that decreased NRG levels are a common characteristic of obesity. In liver and adipose tissue, the increase in NRG expression has protective effects against obesity. However, it is still unknown whether ErbBs expression is altered in this pathology. We hypothesized that high fat diet-induced obesity downregulates ErbB receptors expression in obese mice compared to normal weight mice. Males C57BL/6 mice (n=6-7 for each group) were fed for 12 weeks and divided into: (i) control diet (CD; 10%-kcal fat, 20%-kcal protein, 70%-kcal carbohydrates), and (ii) high fat diet (HFD; 60%-kcal fat, 20%-kcal protein, 20%-kcal carbohydrates). General parameters and ErbBs expression (qPCR, immunohistochemistry and Western blot) were evaluated. We observed a significant increase in final body weight (47%), adipose tissue to body weight ratio (244%) and HOMA-IR (69%), among other parameters, in obese mice. In HFD group significantly decreased ErbB2 (48%) and ErbB3 (66%) mRNA levels in liver (no change in ErbB4), and ErbB2 (43%), ErbB3 (76%) and ErbB4 (35%) in adipose tissue, compared to CD. Furthermore, ErbB2 and ErbB3 protein levels decreased significantly in HFD group compared to the CD in liver. Therefore, our results suggest that HFD-induced obesity significantly decreases ErbBs expression in liver and adipose tissue in this murine model, that may be associated with alterations in the NRG pathway in obese mice.

Las neuregulinas (NRGs) son una familia de proteínas de señalización que se unen a receptores tirosina quinasas de la familia ErbB (ErbB2 a ErbB4), que pueden homo- o heterodimerizar dependiendo de sus características estructurales y del tipo celular. Estudios han propuesto que la disminución de los niveles de NRG es una característica común de la obesidad. En el hígado y el tejido adiposo (TA), el aumento de la expresión de NRG tiene efectos protectores contra la obesidad. Sin embargo, aún se desconoce si la expresión de ErbBs está alterada en esta patología. Nuestra hipótesis es que la obesidad inducida por una dieta alta en grasas (DAG) disminuye la expresión de los ErbB en ratones obesos. Ratones machos C57BL/6 (n=6-7 para c/grupo) fueron alimentados durante 12 semanas y divididos en: (i) dieta control (DC; 10%-kcal grasa, 20%-kcal proteína, 70%-kcal carbohidratos), y (ii) DAG (60%-kcal grasa, 20%-kcal proteína, 20%-kcal carbohidratos). Se evaluaron los parámetros generales y la expresión de ErbBs (qPCR, inmunohistoquímica y Western blot). Observamos un aumento significativo del peso corporal final (47%), de la relación tejido adiposo/peso corporal (244%) y del HOMA-IR (69%), entre otros parámetros, en ratones obesos. En este grupo disminuyó significativamente los niveles de ARNm de ErbB2 (48%) y ErbB3 (66%) en el hígado (sin cambios en ErbB4), y de ErbB2 (43%), ErbB3 (76%) y ErbB4 (35%) en el TA. Además, los niveles de proteína ErbB2 y ErbB3 disminuyeron significativamente, en comparación con el grupo DC en el hígado. Nuestros resultados sugieren que la obesidad inducida por DAG disminuye significativamente la expresión de ErbBs en el hígado y el TA, que puede estar asociado con alteraciones en la vía NRG en ratones obesos.

A convenient and time-saving method for primary culture of mature white adipocytes from mice / 南方医科大学学报

OBJECTIVE@#To establish a simple, low-cost and time-saving method for primary culture of mature white adipocytes from mice.@*METHODS@#Mature white adipocytes were isolated from the epididymis and perirenal area of mice for primary culture using a modified mature adipocyte culture method or the ceiling culture method. The morphology of the cultured mature adipocytes was observed using Oil Red O staining, and the cell viability was assessed with CCK8 method. The expression of PPARγ protein in the cells was detected with Western blotting, and the mRNA expressions of CD36, FAS, CPT1A and FABP4 were detected using RT-qPCR.@*RESULTS@#Oil Red O staining showed a good and uniform morphology of the adipocytes in primary culture using the modified culture method, while the cells cultured using the ceiling culture method exhibited obvious morphological changes. CCK8 assay showed no significant difference in cell viability between freshly isolated mature white adipocytes and the cells obtained with the modified culture method. Western blotting showed that the freshly isolated adipocytes and the cells cultured for 72 h did not differ significantly in the expression levels of PPARγ protein (P=0.759), which was significantly lowered in response to treatment with GW9662 (P < 0.001). GW9662 treatment of the cells upregulated mRNA expressions of CD36 (P < 0.001) and CPT1A (P=0.003) and down-regulated those of FAS (P=0.001) and FABP4 (P < 0.001).@*CONCLUSION@#We established a convenient and time-saving method for primary culture mature white adipocytes from mice to facilitate further functional studies of mature adipocytes.

Gene cloning and sequence analysis of the RPL29 gene and its effect on lipogenesis in goat intramuscular adipocytes / 生物工程学报

The aim of this study was to clone the goat RPL29 gene and analyze its effect on lipogenesis in intramuscular adipocytes. Using Jianzhou big-eared goats as the object, the goat RPL29 gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR), the gene structure and expressed protein sequence were analyzed by bioinformatics, and the mRNA expression levels of RPL29 in various tissues and different differentiation stages of intramuscular adipocytes of goats were detected by quantitative real-time PCR (qRT-PCR). The RPL29 overexpression vector pEGFP-N1-RPL29 constructed by gene recombination was used to transfect into goat intramuscular preadipocytes and induce differentiation. Subsequently, the effect of overexpression of RPL29 on fat droplet accumulation was revealed morphologically by oil red O and Bodipy staining, and changes in the expression levels of genes related to lipid metabolism were detected by qRT-PCR. The results showed that the length of the goat RPL29 was 507 bp, including a coding sequence (CDS) region of 471 bp which encodes 156 amino acid residues. It is a positively charged and stable hydrophilic protein mainly distributed in the nucleus of cells. Tissue expression profiling showed that the expression level of this gene was much higher in subcutaneous adipose tissue and inter-abdominal adipose tissue of goats than in other tissues (P < 0.05). The temporal expression profile showed that the gene was expressed at the highest level at 84 h of differentiation in goat intramuscular adipocytes, which was highly significantly higher than that in the undifferentiated period (P < 0.01). Overexpression of RPL29 promoted lipid accumulation in intramuscular adipocytes, and the optical density values of oil red O staining were significantly increased (P < 0.05). In addition, overexpression of RPL29 was followed by a highly significant increase in ATGL and ACC gene expression (P < 0.01) and a significant increase in FASN gene expression (P < 0.05). In conclusion, the goat RPL29 may promote intra-muscular adipocyte deposition in goats by up-regulating FASN, ACC and ATGL.

Hibernoma, tumor benigno infrecuente

El hibernoma es un tumor limítrofe, benigno de comportamiento agresivo, de tejidos blandos, raro en frecuencia, que se origina a partir de los adipocitos pardos residuales, afectando principalmente a mujeres entre 10 a 20 años. Clínicamente se presenta como tumoración, más frecuente en los muslos, de crecimiento lento, móvil, firme e indolora. En los estudios de imagen, puede resultar difícil realizar un diagnóstico diferencial por su similitud con neoplasias malignas. El tratamiento ideal es la resección quirúrgica con márgenes amplios para evitar cualquier tipo de recurrencia local. Se presenta el caso de un hibernoma típico, manejo y tratamiento quirúrgico realizado.

Hibernoma is a borderline tumor, benign with aggressive behavior, of soft tissues, rare in frequency, which originates from residual brown adipocytes, mainly affecting women between 10 and 20 years of age. Clinically it presents as a slow-growing, mobile, firm and painless tumor, more frequent on the thighs. In imaging studies, it can be difficult to make a differential diagnosis due to its similarity to malignant neoplasms. The ideal treatment is surgical resection with wide margins to avoid any type of local recurrence. The case of a typical hibernoma, management and surgical treatment performed is presented.

Bone Marrow Adipocytes Promote the Survival of Multiple Myeloma Cells and Up-Regulate Their Chemoresistance / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of adipocytes in the bone marrow microenvironment of patients with multiple myeloma (MM) on the pathogenesis of MM.@*METHODS@#Bone marrow adipocytes (BMA) in bone marrow smears of health donors (HD) and newly diagnosed MM (ND-MM) patients were evaluated with oil red O staining. The mesenchymal stem cells (MSC) from HD and ND-MM patients were isolated, and in vitro co-culture assay was used to explore the effects of MM cells on the adipogenic differentiation of MSC and the role of BMA in the survival and drug resistance of MM cells. The expression of adipogenic/osteogenic differentiation-related genes PPAR-γ, DLK1, DGAT1, FABP4, FASN and ALP both in MSC and MSC-derived adipocytes was determined with real-time quantitative PCR. The Western blot was employed to detect the expression levels of IL-6, IL-10, SDF-1α, TNF-α and IGF-1 in the supernatant with or without PPAR-γ inhibitor.@*RESULTS@#The results of oil red O staining of bone marrow smears showed that BMA increased significantly in patients of ND-MM compared with the normal control group, and the BMA content was related to the disease status. The content of BMA decreased in the patients with effective chemotherapy. MM cells up-regulated the expression of MSC adipogenic differentiation-related genes PPAR-γ, DLK1, DGAT1, FABP4 and FASN, but the expression of osteogenic differentiation-related gene ALP was significantly down-regulated. This means that the direct consequence of the interaction between MM cells and MSC in the bone marrow microenvironment is to promote the differentiation of MSC into adipocytes at the expense of osteoblasts, and the cytokines detected in supernatant changed. PPAR-γ inhibitor G3335 could partially reverse the release of cytokines by BMA. Those results confirmed that BMA regulated the release of cytokines via PPAR-γ signal, and PPAR-γ inhibitor G3335 could distort PPAR-γ mediated BMA maturation and cytokines release. The increased BMA and related cytokines effectively promoted the proliferation, migration and drug resistance of MM cells.@*CONCLUSION@#The BMA and its associated cytokines are the promoting factors in the survival, proliferation and migration of MM cells. BMA can protect MM cells from drug-induced apoptosis and plays an important role in MM treatment failure and disease progression.

Effect of autophagic core protein Atg101 on adipocyte senescence / 中华内分泌代谢杂志

Objective:To investigate the effect of autophagy related gene Atg101 on white adipocyte senescence.Methods:An Atg101 knockdown model of 3T3-L1 mature adipocytes was constructed to probe the effect of Atg101 on autophagy-related proteins LC3 and p62 protein. The RNA-seq database of human subcutaneous adipose tissue was constructed and analyzed, and the co-expressed gene set was predicted based on the pearson correlation coefficient( R2>0.4, P<0.05) between FPKM values of Atg101 and other gene, followed by KEGG and Reactome enrichment analysis. Young mouse(8 weeks old) and old mouse(18 months old) models were established, and the expression levels of Atg101 in inguinal white adipose tissue and epididymal white adipose tissue were detected by quantitative real-time PCR(RT-qPCR) and Western blot. Furthermore, the differences in white adipocyte senescence-associated secretory phenotype(SASP), cell cycle and mitochondrial homeostasis-related genes were detected by RNA-seq, Western blot, and RT-qPCR to analyze the effects of Atg101 silencing on adipocyte senescence. Results:The autophagy-related protein LC3-Ⅱ expression was significantly decreased and p62 protein was induced after Atg101 was knockdowned in 3T3-L1 adipocytes, suggesting impaired cell autophagy. KEGG enrichment analysis revealed that Atg101 co-expressed gene set was mainly enriched in autophagy and senescence-related pathways; Reactome enrichment analysis revealed that this gene set was associated with multiple cell cycle signaling pathways. RT-qPCR and Western blot confirmed that both mRNA and protein levels of Atg101 were down-regulated in inguinal white adipose tissue of aging mice, and protein levels in epididymal white adipose tissue were also significantly reduced. Finally, it was further confirmed that SASP-related genes were induced after Atg101 knockdown in white adipocytes, and cell cycle-specific gene expression was restricted and cytokine-dependent protein kinase inhibitors p16 and p21 expressions were significantly increased, while mitochondrial homeostasis regulatory genes were also suppressed.Conclusions:Knockdown of Atg101 may regulate white adipocyte senescence by inhibiting autophagic activity, presenting impaired mitochondrial homeostasis.

Influencia de ácidos grasos en parámetros sanguíneos y adipogénesis. Estudio experimental en pollos eclosionados / Influence of fatty acids on blood parameters and formation of adipocytes at early age

Introducción: La alimentación en la primera infancia influencia la instauración del tejido adiposo y el desarrollo de diversas patologías en la edad adulta. Objetivo: Evaluar la influencia del consumo de tres fuentes de ácidos grasos sobre parámetros sanguíneos y tejido adiposo en pollos recién eclosionados. Materiales y métodos: Se utilizaron 76 pollitos Cobb 500 distribuidos aleatoriamente en cuatro tratamientos, que fueron alimentados durante siete días con una de las cuatro dietas (T1: 97% Dieta basal (DB); T2: DB +3% de manteca vegetal parcialmente hidrogenada; T3: DB +3% de aceite de quinua y T4: DB +3% de aceite de pescado). Al finalizar, se evaluó en sangre glucosa, colesterol, triglicéridos y tamaño de adipocitos del tejido adiposo subcutáneo y visceral. Se aplicó ANOVA considerando 0,05 de significancia y en el caso de variables no distribuidas normalmente, se aplicó el test no paramétrico de Kruskal-Wallis mediante el programa R-Studio. Resultados: Se obtuvieron diferencias significativas con disminución de los niveles de glucosa y colesterol en animales suplementados con elevada proporción de aceites insaturados (T3 y T4) en comparación a T2. Los tratamientos T3 y T4 promovieron una formación hiperplásica de adipocitos, diferenciándose significativamente de T2, que promovió la hipertrofia en dichas células, esta respuesta fue similar en ambos depósitos subcutáneos. Conclusiones: El consumo de aceite de quinua y aceite de pescado promueve la formación de tejido adiposo saludable, y reducen los niveles de glucosa y colesterol. Contrariamente el consumo de manteca vegetal propicia la hipertrofia de adipocitos de gran tamaño e incrementa los parámetros bioquímicos evaluados(AU)

Introduction: Feeding in early childhood influences the establishment of adipose tissue, and therefore also in the development of various pathologies in adulthood. Objective: To evaluate the influence of the consumption of three sources of fatty acids on blood parameters and adipose tissue at an early age. Materials and methods: 76 Cobb 500 chicks randomly distributed in four treatments were used, who were fed for seven days with one of the four diets (T1: Basal diet; T2: DB +1.0% vegetable shortening partially hydrogenated; T3: DB +1.0% quinoa oil and DB +1.0% fish oil) until the seventh day of life. At the end, glucose, cholesterol, triglycerides and adipocyte size of the subcutaneous and visceral adipose tissue were evaluated. A completely randomized design with ANOVA considering 0.05 significance was applied and in the case of non-normally distributed variables, the non-parametric Kruskal-Wallis test was applied using the R-Studio program. Results: Significant differences were obtained with a decrease in glucose and cholesterol levels in animals supplemented with unsaturated oils (T3 and T4) compared to T2. Regarding the size of adipocytes, treatments T3 and T4 promoted a hyperplastic formation of adipocytes, differing significantly from T2, which promoted hypertrophy in these cells, this response was similar in both subcutaneous deposits. Conclusions: The consumption of quinoa oil and fish oil promote the formation of healthy adipose tissue, in addition to reducing glucose and cholesterol levels. In contrast, the consumption of vegetable shortening favors the hypertrophy of large adipocytes and increases the biochemical parameters evaluated(AU)

Inovação em cânula de lipoaspiração para coleta de enxerto de gordura para uso em reconstrução mamária / Innovation in liposuction cannula to harvest fat graft to use in breast reconstruction

Introdução: A evolução da técnica de lipoaspiração e da enxertia de gordura associada a segurança oncológica permitiram utilizar a gordura como preenchedor autólogo em pacientes com neoplasia de mama que eram submetidas a reconstrução mamária. O objetivo é apresentar uma inovação no instrumento de coleta de gordura que será utilizada como enxerto, para uso em reconstrução mamária. Métodos: Foi realizada a busca de anterioridade nos bancos de dados internacionais e nacional e a confecção de um protótipo de cânula de coleta de gordura de 35cm de comprimento, com diâmetro do tubo de 4mm, com 10 furos redondos na sua extremidade distal com 2mm de diâmetro. Foi realizado um experimento de aspiração de substância viscosa comparando o protótipo com dois modelos de cânulas com desenho padrão da indústria, de três furos tipo Mercedes e de cinco furos tipo Pitanguy. Resultados: Foram encontrados quatro patentes de alta relevância e uma patente de média relevância, que diferem do modelo de utilidade proposto quando se comparam o tipo de desenho da extremidade distal das cânulas estudadas e a função dos instrumentos identificados. Quando comparou-se a eficiência do protótipo, a cânula de cinco furos foi a mais eficiente na aspiração da substância viscosa, e não houve diferença estatística na velocidade de aspiração entre o protótipo e a cânula de três furos tipo Mercedes. Conclusão: A inovação apresentada para a coleta de enxerto de gordura para uso em reconstrução mamária apresentou a mesma eficiência que a cânula Mercedes de três furos neste modelo experimental.

Introduction: The evolution of liposuction and fat graft technique with the oncologic safety allowed to use the fat as autologous filler in patients with breast cancer and submitted to breast reconstruction. The objective is to introduce an innovation in the instrument used to harvest fat grafts for breast reconstruction. Methods: Search of anteriority and trademarks were performed at international and national databases, and a prototype was built as a fat harvest instrument with 35cm in length, 4cm in diameter, with 10 holes at the distal extremity, with 2mm diameter each. It was experimented with a harvest of a viscous substance, comparing the prototype with two regular industry cannulas types: three holes Mercedes' type and five roles Pitanguy's type. Results: Four high-relevance patents and one medium-relevance patent were found, which differ from the proposed utility model when comparing the type of design of the distal end of the studied cannulas and the function of the instruments identified. When the prototype's efficiency was compared with the other cannulas, the cannula with five holes in Pitanguy's type was the most efficient, and it was no statistical difference between the prototype and the cannula with three roles in Mercedes' type. Conclusion: The innovation presented to harvest fat graft for breast reconstruction had the same efficiency in harvesting the viscose substance as the cannula Mercedes type with three holes in this experimental model.

Lipoleiomioma uterino

El lipoleiomioma es una neoplasia uterina benigna poco frecuente cuya incidencia varía entre 0,03% y 0,2%. Este tumor es considerado una variante benigna de los leiomiomas uterinos típicos. Está formado por una proporción variable de adipocitos maduros y células musculares lisas. La etiología puede estar relacionada con la deficiencia de estrógenos que se produce después de la transición menopáusica; generalmente aparece en mujeres obesas perimenopáusicas o menopáusicas. La sintomatología es inespecífica y la mayoría es diagnosticada de forma incidental. Se presenta un caso de lipoleiomioma uterino en paciente de 45 años quien consultó por presentar dolor abdominal. La ecografía mostró tumor en pared anterior de un útero homogéneo y bien definido. Durante la laparotomía se encontró tumor amarillento y de textura blanda. Se realizó histerectomía total más ooforosalpingectomía. El diagnóstico anatomopatológico fue de lipoleiomioma uterino.

Lipoleiomyoma is a rare benign uterine neoplasm whose incidence varies between 0.03%-0.2%. This tumor is considered a benign variant of typical uterine leiomyomas. It consists of a variable proportion of mature adipocytes and smooth muscle cells. The etiology may be related to estrogen deficiency occurring after the menopausal transition; it usually appears in obese perimenopausal or menopausal women. The symptomatology is nonspecific, and most are diagnosed incidentally. We present a case of uterine lipoleiomyoma in a 45-year-old patient who consulted for abdominal pain. Ultrasonography showed a tumor in the anterior wall of a homogeneous and well-defined uterus. During laparotomy, a yellowish tumor with a soft texture was found. Total hysterectomy plus oophorosalpingectomy was performed. The anatomopathologic diagnosis was uterine lipoleiomyoma.

Espectros clínicos y fisiopatológicos del síndrome de Bernardinelli Seip / Clinical and pathophysiological spectra of Bernardinelli Seip syndrome

Introducción: La lipodistrofia congénita de Berardinelli-Seip es un síndrome genético autosómico recesivo, caracterizado por la ausencia generalizada del tejido adiposo, el déficit de leptina y las alteraciones metabólicas incluidas la resistencia a la insulina, la esteatohepatitis y la hipertrigliceridemia. Objetivo: Definir los diferentes espectros clínicos y fisiopatológicos del síndrome y su relación con el fenotipo definiendo las estrategias terapéuticas actuales. Métodos: Se realizó una búsqueda bibliográfica no sistemática en las bases de datos Science Direct, EMBASE, LILACS, Redalyc, SciELO y PubMed. Los criterios de inclusión fueron publicaciones en inglés, portugués o español, en las que el título y las palabras clave, abordaban el tema planteado con una vigencia de 10 años. Se obtuvieron 50 artículos relacionados con el síndrome, de los cuales 30 fueron seleccionados para su revisión. Conclusiones: El diagnóstico de la enfermedad es principalmente clínico. Se establece en presencia de tres criterios mayores o la combinación de dos mayores con dos menores y/o por la identificación de variantes patogénicas por medio del estudio genético y molecular. La dieta y el ejercicio conjuntamente con la administración de la metreleptina son pilares fundamentales en el manejo de estos pacientes. El reconocimiento temprano del síndrome es esencial para prevenir las complicaciones, y brindar asesoría genética y reproductiva a los pacientes y familiares(AU)

Introduction: Berardinelli-Seip congenital lipodystrophy is an autosomal recessive genetic syndrome, characterized by the general absence of adipose tissue, leptin deficiency and metabolic alterations including insulin resistance, steatohepatitis and hypertriglyceridemia. Objective: To present the different clinical and pathophysiological spectra of the syndrome, its relationship with the phenotype, defining the current therapeutic strategies. Methods: A non-systematic bibliographic search was carried out in Science Direct, EMBASE, LILACS, Redalyc, SciELO and PubMed databases. The inclusion criteria were publications in English, Portuguese and Spanish, in which the title and keywords included information pertinent to the stated objective with a periodicity of 10 years, 50 articles were retrieved, and 30 of them were selected. Conclusions: The diagnosis of the disease is mainly clinical. It is established in the presence of three major criteria or the combination of two major and two minor criteria and/or by the identification of pathogenic variants through genetic and molecular studies. Diet and exercise together with the administration of metreleptin are fundamental pillars in the management of these patients. Early recognition of the syndrome is essential to prevent complications, allowing genetic and reproductive counseling to be provided to patients and their families(AU)

Hipertrofia y resistencia a la insulina en un modelo in vitro de obesidad y DMT2 inducida por alta glucosa e insulina / Hypertrophy and insulin resistance in an in vitro model of obesity and T2DM induced by high glucose and insulin

Resumen Introducción: La obesidad es considerada un factor de riesgo para desarrollar resistencia a la insulina. La expansión del tejido adiposo se ha relacionado con el aumento de la producción de citoquinas proinflamatorias que, junto a los ácidos grasos son responsables, al menos en parte, del desarrollo de la resistencia a la insulina y esta a su vez, facilita el desarrollo de diabetes mellitus tipo 2 (DMT2). Objetivo: El propósito de este estudio fue realizar y caracterizar un modelo in vitro de obesidad empleando concentraciones altas de glucosa e insulina en una línea de células adipocitarias. Métodos: Se indujo modelo de hipertrofia celular realizando un estímulo en adipocitos maduros con una concentración de glucosa (450 mg/dL) e insulina (106 pmol/L) (modelo HGHI). Tras estímulo se realizaron ensayos de viabilidad celular, diámetro celular, movilización de lípidos y marcadores de señalización de insulina. Resultados: Tras el tratamiento con HGHI, se evidencia hipertrofia adipocitaria, incremento en la acumulación de lípidos, reducción de la ruptura de éstos, alteración de la señalización de insulina y tendencia a modificación de proteínas de marcadores de estrés de retículo y estrés oxidativo. Conclusión: Estos resultados demuestran la validez del modelo in vitro que simula al menos en parte la obesidad asociada a insulino resistencia, siendo una herramienta útil para estudiar los mecanismos de susceptibilidad a obesidad y resistencia a la insulina inducida in vitro con diferentes moléculas.

Abstract Introduction: Obesity is considered a risk factor for developing insulin resistance. The increase in adipose tissue has been related to the increase in the production of pro-inflammatory cytokines, which together with fatty acids are responsible, at least in part, for the development of insulin resistance, and this in turn facilitates the development of T2 diabetes mellitus type 2 (DMT2). Objective: The purpose of this study was to perform and characterize an in vitro model of obesity using high concentrations of glucose and insulin on an adipocyte cell line. Methods: A cell hypertrophy model was induced by stimulating mature adipocytes with a concentration of glucose (450 mg/dL) and insulin (106 pmol/L) (HGHI model). The cell viability, cell diameter, lipid mobilization and insulin signalling markers were evaluated. Results: After HGHI treatment, adipocytes show hypertrophy, increase in lipid accumulation, reduction of lipid breakdown, alteration of insulin signalling, a tendency to modify proteins of reticulum stress markers and, oxidative stress. Conclusion: These results demonstrate a new in vitro model that simulates, at least in part, obesity associated with insulin resistance being a useful tool to study the mechanisms of susceptibility to obesity and insulin resistance induced in vitro by different molecules.

Sports medicine to reduce obesity body fat rate and promote health research / Medicina do esporte na redução da taxa de gordura corporal na obesidade e promoção de pesquisas na saúde / Medicina del deporte en la reducción de la tasa de grasa corporal en obesidad y promoción de investigaciones en la salud

ABSTRACT Introduction: Obesity is a chronic disease with excessive accumulation of body fat, exceeding the normal health level and affecting the physiological functioning of the body. Objective: To establish an obesity evaluation standard closely related to health based on sports medicine indexes, and to provide a new theory and method for the formulation of an obesity evaluation standard in China. Methods: Height, weight, skin fold thickness (shoulder, arm) and medical indexes (SBP, DBP, TC, TG, HDL, INS) of 108 female subjects in a city were tested by means of anthropometry, experiment, and mathematical statistics. All the selected medical indicators were evaluated according to clinical medical standards. Results: the body mass index of 108 women was positively correlated with such medical indexes as systolic blood pressure, diastolic blood pressure, triglycerides, high density lipoprotein cholesterol, insulin and so on (P<0.05~0.01). Conclusions: The correlation between medical indexes and BMI is high, and the abnormal rate increases with the increase in the level of obesity. Clinical indicators can be used as an important scientific basis for the establishment of obesity evaluation criteria. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: A obesidade é uma doença crônica em que há acumulo excessivo de gordura corporal, excedendo o nível saudável normal e afetando o funcionamento fisiológico do corpo. Objetivo: Estabelecer uma avaliação padrão de obesidade estritamente relacionada a saúde com base nos índices de medicina do esporte, e fornecer uma nova teoria e novo método para a formulação de uma avaliação padrão de obesidade na China. Métodos: Altura, peso, espessura de dobras cutâneas (ombro, braço) e índices médicos (PSS, PSD, TC, HDL, INS) de 108 sujeitos do sexo feminino, moradoras de uma cidade, foram tomados por meio de antropometria, experimentos e estatísticas matemáticas. Todos os indicadores médicos selecionados foram avaliados de acordo com padrões médicos clínicos. Resultados: o índice de massa corporal de 108 mulheres se correlaciona com índices médicos como pressão sanguínea sistólica, pressão sanguínea diastólica, triglicerídeos, alta densidade de colesterol lipoproteico, insulina e assim por diante (P<0.05~0.01). Conclusões: A correlação entre índices médicos e o IMC é alta e a taxa anormal aumenta com o aumento no grau de obesidade. Indicadores clínicos podem ser usados como base cientifica importante para o estabelecimento de critérios de avaliação de obesidade. Nível de evidência II; Estudos terapêuticos - investigação de resultados de tratamento.

RESUMEN Introducción: La obesidad es una enfermedad crónica en que hay acúmulo excesivo de grasa corporal, excediendo el nivel saludable normal y afectando el funcionamiento fisiológico del cuerpo. Objetivo: Establecer una evaluación estándar de obesidad estrictamente relacionada a la salud con base en los índices de medicina del deporte, y brindar una nueva teoría y método para la formulación de una evaluación estándar de obesidad en China. Métodos: Altura, peso, espesor de pliegues cutáneos (hombro, brazo) e índices médicos (PSS, PSD, TC, HDL, INS) de 108 sujetos del sexo femenino, residentes de una ciudad, se tomaron por medio de antropometría, experimentos y estadísticas matemáticas. Todos los indicadores médicos seleccionados se evaluaron de acuerdo a patrones médicos clínicos. Resultados: El índice de masa corporal de 108 mujeres se correlaciona a índices médicos como presión sanguínea sistólica, presión sanguínea diastólica, triglicéridos, colesterol de lipoproteínas de alta densidad, insulina y así sucesivamente (P<0.05~0.01). Conclusiones: La correlación entre índices médicos y el IMC es alta y la tasa anormal aumenta con el aumento en el grado de obesidad. Indicadores clínicos pueden usarse como base científica importante para el establecimiento de criterios de evaluación de obesidad. Nivel de evidencia II; Estudios terapéuticos - investigación de resultados de tratamiento.

Relação entre resposta imune inata do receptor toll-like-4 (TLR-4) e o processo fisiopatológico da cardiomiopatia da obesidade / Relationship between innate immune response toll-like receptor 4 (TLR-4) and the pathophysiological process of obesity cardiomyopathy

Resumo Fundamento A obesidade é uma condição inflamatória crônica de baixo grau relacionada a distúrbios cardíacos. No entanto, o mecanismo responsável pela inflamação cardíaca relacionada à obesidade não é claro. O receptor do tipo toll 4 (TLR-4) pertence a um receptor da família das transmembranas, responsável pela resposta imune, cuja ativação estimula a produção de citocinas pró-inflamatórias. Objetivo Testar se a ativação do receptor TLR-4 participa do processo de cardiomiopatia da obesidade, devido à produção de citocinas por meio da ativação do NF-ĸB. Métodos Ratos Wistar machos foram randomizados em dois grupos: o grupo controle (C, n = 8 animais) que recebeu dieta padrão/água e o grupo obeso (OB, n = 8 animais) que foi alimentado com dieta rica em açúcar e gordura e água mais 25% de sacarose por 30 semanas. Análise nutricional: peso corporal, índice de adiposidade, alimentos, água e ingestão calórica. Análise de distúrbios relacionados à obesidade: glicose plasmática, ácido úrico e triglicerídeos, HOMA-IR, pressão arterial sistólica, TNF-α no tecido adiposo. A análise cardíaca incluiu: expressão das proteínas TLR-4 e NF-ĸB, níveis de TNF-α e IL-6. Comparação pelo teste t de Student não pareado ou teste de Mann-Whitney com um valor de p <0,05 como estatisticamente significativo. Resultados O grupo OB apresentou obesidade, glicose elevada, triglicerídeos, ácido úrico, HOMA, pressão arterial sistólica e TNF-α no tecido adiposo. O grupo OB apresentou remodelação cardíaca e disfunção diastólica. A expressão de TLR-4 e NF-ĸB e os níveis de citocinas foram maiores em OB. Conclusão Nossos achados concluem que, em uma condição obesogênica, a inflamação derivada da ativação do TLR-4 cardíaco pode ser um mecanismo capaz de levar à remodelação e disfunção cardíaca.

Abstract Background Obesity is a chronic low-grade inflammation condition related to cardiac disorders. However, the mechanism responsible for obesity-related cardiac inflammation is unclear. The toll-like receptor 4 (TLR-4) belongs to a receptor of the transmembrane family responsible for the immune response whose activation stimulates the production of proinflammatory cytokines. Objective To test whether the activation of the TLR-4 receptor participates in the obesity cardiomyopathy process, due to cytokine production through NF-ĸB activation. Methods Male Wistar rats were randomized into two groups: the control group (C, n= 8 animals) that received standard diet/water and the obese group (OB, n= 8 animals) that were fed a high sugar-fat diet and water plus 25% of sucrose for 30 weeks. Nutritional analysis: body weight, adiposity index, food, water, and caloric intake. Obesity-related disorders analysis: plasma glucose, uric acid and triglycerides, HOMA-IR, systolic blood pressure, TNF-α in adipose tissue. Cardiac analysis included: TLR-4 and NF-ĸB protein expression, TNF-α and IL-6 levels. Comparison by unpaired Student's t-test or Mann- Whitney test with a p-value < 0.05 as statistically significant. Results The OB group showed obesity, high glucose, triglycerides, uric acid, HOMA, systolic blood pressure, and TNF-α in adipose tissue. OB group presented cardiac remodeling and diastolic dysfunction. TLR-4 and NF-ĸB expression and cytokine levels were higher in OB. Conclusion Our findings conclude that, in an obesogenic condition, the inflammation derived from cardiac TLR-4 activation can be a mechanism able to lead to remodeling and cardiac dysfunction.

Viabilidade das células-tronco mesenquimais do tecido adiposo proveniente de lipoaspirado humano / Viability of mesenchymal stem cells of adipose tissue from human liposuction

Introdução: A lipoenxertia é uma alternativa com importante aplicabilidade para reconstrução de mama e/ou correções de assimetrias decorrentes do tratamento oncológico. Esta técnica consiste na transferência de gordura autóloga, cujo estroma contém células-tronco derivadas do tecido adiposo que tem capacidade de diferenciar-se em toda a linhagem mesodermal. Para o preparo do tecido adiposo, Coleman fundamentou a centrifugação, de material aspirado por seringa, em 3000 rotações por minuto (rpm) durante 3 minutos. Contudo, estudos questionam se velocidades menores de centrifugação poderiam ser menos deletérias para viabilidade celular. Métodos: Foi realizado um estudo experimental, onde foram avaliadas as células adiposas de seis pacientes; a partir de 60mL de lipoaspirado de cada um. A amostra coletada foi fracionada em quatro tubos, e submetidos a diferentes protocolos, decantação e centrifugação nas velocidades 500, 1000 e 3000rpm por 3 minutos. Após as amostras foram processadas com colagenase IA por 30 min, submetidas ao cultivo celular por 24 horas e realizado a análise da viabilidade celular. Os resultados foram tabulados e analisados pelo teste ANOVA utilizando os programas Graphpad Prism 6.0® e SAS®. Resultados: A viabilidade celular foi maior na amostra celular centrifugada a 3000rpm e menor na amostra decantada. A coloração com Giemsa indicou manutenção da morfologia celular entre as amostras. Conclusão: As células centrifugadas na velocidade de 3000rpm apresentaram maior viabilidade celular. A centrifugação foi efetiva na compactação do tecido e eliminação de resíduos indesejados (sangue e óleo residual).

Introduction: Lipografting is an alternative with important applicability for breast reconstruction and/or corrections of asymmetries resulting from cancer treatment. This technique consists of autologous fat transfer, whose stroma contains stem cells derived from adipose tissue that can differentiate itself throughout the mesodermal lineage. For adipose tissue preparation, Coleman-based centrifugation of syringe-aspirated material at 3000 revolutions per minute (rpm) for 3 minutes. However, studies question whether lower centrifugation speeds could be less harmful to cell viability. Methods: An experimental study was conducted to evaluate the adipose cells of six patients; from 60mL of liposuction of each one. The sample collected was fractionated into four tubes and submitted to different protocols, decanting and centrifugation at speeds 500, 1000, and 3000rpm for 3 minutes. Afterward, the samples were processed with collagenase IA for 30 min, submitted to cell culture for 24 hours, and a cell viability analysis. The results were tabulated and analyzed by the ANOVA test using the Graphpad Prism 6.0® and SAS®. Results: Cell viability was higher in the cell sample centrifuged at 3000rpm and lower in the decanted sample. Giemsa staining indicated maintenance of cell morphology on the samples. Conclusion: Centrifuged cells at a speed of 3000rpm showed higher cell viability. Centrifugation was effective in compacting tissue and eliminating unwanted waste (blood and residual oil).

An in vitro model mimicking the complement system to favor directed phagocytosis of unwanted cells

BACKGROUND: Opsonization, is the molecular mechanism by which target molecules promote interactions with phagocyte cell surface receptors to remove unwanted cells by induced phagocytosis. We designed an in vitro system to demonstrate that this procedure could be driven to eliminate adipocytes, using peptides mimicking regions of the complement protein C3b to promote opsonization and enhance phagocytosis. Two cell lines were used: (1) THP-1 monocytes differentiated to macrophages, expressing the C3b opsonin receptor CR1 in charge of the removal of unwanted coated complexes; (2) 3T3-L1 fibroblasts differentiated to adipocytes, expressing AQP7, to evaluate the potential of peptides to stimulate opsonization. (3) A co-culture of the two cell lines to demonstrate that phagocytosis could be driven to cell withdrawal with high efficiency and specificity. RESULTS: An array of peptides were designed and chemically synthesized p3691 and p3931 joined bound to the CR1 receptor activating phagocytosis (p < 0.033) while p3727 joined the AQP7 protein (p < 0.001) suggesting that opsonization of adipocytes could occur. In the co-culture system p3980 and p3981 increased lipid uptake to 91.2% and 89.0%, respectively, as an indicator of potential adipocyte phagocytosis. CONCLUSIONS: This in vitro model could help understand the receptor­ligand interaction in the withdrawal of unwanted macromolecules in vivo. The adipocyte-phagocytosis discussed may help to control obesity, since peptides of C3b stimulated the CR1 receptor, promoting opsonisation and phagocytosis of lipidcontaining structures, and recognition of AQP7 in the differentiated adipocytes, favored the phagocytic activity of macrophages, robustly supported by the co-culture strategy.

Effect of Daizongfang on Insulin Resistance of 3T3-L1 Adipocytes Induced by Different Methods / 中国实验方剂学杂志

Objective:To investigate the effects of Daizongfang （DZF） on insulin resistance （IR） of adipocytes induced by different methods. Method:The cocktail induction method was adopted to induce the differentiation and maturity of 3T3-L1 preadipocytes. An IR model in mature adipocytes was established by the induction of palmitic acid （PA）， high-concentration glucose （HG）， and dexamethasone （DEX）. DZF extracts at different concentrations （2.0， 0.5， 0.1 g·L^-1） intervened for 24 hours. A model group， a rosiglitazone （RSG） group， and a blank control group were set up at the same time. The glucose concentration in the culture supernatant was measured by the glucose oxidase-peroxidase （GOD-POD） method. Glucose consumptions under basic conditions （G_Basic） and insulin stimulation （G_Ins） were calculated to evaluate the insulin sensitivity index （ISI）. The mRNA expression of glucose transporter 4 （GLUT4） was detected by the real-time polymerase chain reaction （PCR）. Result:Compared with the model group， the DZF （2.0 g·L^-1） showed increased G_Basic， G_Ins， and ISI in three IR models （<italic>P</italic><0.05， <italic>P</italic><0.01）. In addition， for the PA-induced IR model， G_Basic and G_Ins in the DZF （0.5 g·L^-1） group were elevated （<italic>P</italic><0.01）， and G_Basic， G_Ins， and ISI in the RSG group increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. For the HG-induced IR model， G_Ins and ISI increased in the DZF （0.5 g·L^-1） group （<italic>P</italic><0.05）， and G_Basic， G_Ins， and ISI were elevated in the RSG group （<italic>P</italic><0.01）. For the DEX-induced IR model， G_Ins and ISI increased in the RSG group （<italic>P</italic><0.01）. In the three models， there were differences among groups with different doses. G_Basic， G_Ins， and ISI in the high-dose DZF group increased in varying degrees compared with those in the medium- and low-dose DZF groups （<italic>P</italic><0.05）. In the three models， the DZF （2.0 g·L^-1） group and the RSG group both increased GLUT4 mRNA expression （<italic>P</italic><0.05）. Conclusion:DZF can reduce IR of adipocytes induced by HG， DEX， or PA in a dose-dependent manner and increase glucose uptake in an insulin-independent manner， which may be related to the increase in GLUT4 expression.

Transcriptome profiling reveals differential expression of genes potentially involved in muscle and adipose tissue development of cattle

BACKGROUND: To identify differentially expressed genes (DEGs) between muscle and adipose in cattle, we analyzed the data from the RNA sequencing of three Angus×Qinchuan crossbred cattle. RESULTS: Searched the Gene Expression Omnibus (GEO) for a microarray dataset of Yan yellow cattle, GSE49992. After the DEGs were identified, we used STRING and Cytoscape to construct a protein­protein interaction (PPI) network, subsequently analyzing the major modules of key genes. In total, 340 DEGs were discovered, including 21 hub genes, which were mainly enriched in muscle contraction, skeletal muscle contraction, troponin complex, lipid particle, Z disc, tropomyosin binding, and actin filament binding. CONCLUSIONS: In summary, these genes can be regarded as candidate biomarkers for the regulation of muscle and adipose development.

Adiponectin and Body Mass Index in Type 2 Diabetes Mellitus

Introduction: Foot Ulcer is the most frequent complicationof Diabetes Mellitus related to lower extremities with highmorbidity. Infection is an important component of foot ulcer.The present study was undertaken to study the clinical andmicrobiological profile of diabetic foot ulcer in a tertiary carecenter.Material and Methods: It is a prospective observational studywhich has been conducted at VIMSAR, Burla from November2017 to November 2019. Consecutive patients of Diabeticfoot ulcers were included in this study. After admissiondetailed history and biochemical investigations were donein all cases. Nerve Conduction study, doppler of lower limbwere performed to assess peripheral neuropathy and vasculardisease. The ulcers were graded according to Wagner’sstaging. Swabs were taken for culture and sensitivity.Results: Out of 1234 patients of DM admitted to the hospital,116 (9.4%) patients had foot ulcer. There were 87 (75.0%)males and 29 (25.0%) females with M:F ratio of 3:1. 95(81.9%) patients were from rural and 21 (18.1%) from urbanarea. Majority (n=44, 38.0%) of patients belonged to 51-60years of age and only 3 (2.0%) were within 21 to 30 years.Type-1 and -2 DM was found in 4 (3.0%) and 112 (97.0%)patients and the duration of DM was given. Grade-2 ulcerwas found in 62 (53.5%) cases. Risk factors like peripheralneuropathy, retinopathy and nephropathy were present in(n=65, 56.1%), 24 (20.7%) and 35 (30.1%) cases respectively.Early signs of ulcerative lesion like abundant callus, crackfoot, blistering was present in 28 (24.1%), 54 (46.6%),10(8.6%) cases respectively.Conclusion: This study showed that foot ulcer is a commonchronic complication of DM mostly found in elderly patientswith poor glycemic control from rural areas. S.aureus andP.aeruginosa are the common organisms detected from theulcer.

Effect of protease inhibitor from Agaricus bisporus on glucose uptake and oxidative stress in 3T3-L1 adipocytes

Objective: To explore the effect of the protease inhibitor from Agaricus bisporus (J.E. Lange) Imbach (AbPI) on glucose uptake and oxidative stress in 3T3-L1 adipocytes. Methods: Adipocytes were differentiated and stained with Oil-Red-O staining to confirm adipogenesis. The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay, intracellular reactive oxygen species generation through flow cytometry, and morphologically through confocal microscopy using propidium iodide, 4,6-diamino-2-phenylindol dihydrochloride, and 2',7'-dichlorofluorescein diacetate dyes. The uptake of fluorescent glucose analog, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy. Results: MTT assay showed that the cell survival rate was (28.00±3.00)％, (92.33±2.60)％, and (71.34±2.10)％ in the presence of 2 mM H2O2, AbPI alone, and AbPI and H2O2 both, respectively, in comparison to the control. Oil-Red-O staining indicated that AbPI enhanced adipogenesis. AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone, and showed insulin-sensitizing effect in the presence of insulin, but failed to stimulate the uptake in the absence of insulin. Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon AbPI treatment. Confocal microscopy showed that the damaged cell population rose to 3.50％, 117.84％, and 261.50％ in the presence of AbPI alone, AbPI with H2O2, and H2O2 alone, respectively. Conclusions: The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.